Additivities for Soluble Recombinant Protein Expression in Cytoplasm of Escherichia coli

D. L. Atroshenko, Egor P. Sergeev, Diana I. Golovina, A. Pometun
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Abstract

Recombinant protein expression in Escherichia coli is a fundamental technique in molecular biology and biotechnology. This review provides a comprehensive overview of various additivities to enhance the expression levels of soluble recombinant proteins in E. coli. The discussion encompasses five key aspects. Inducer Optimization: strategies for optimizing the inducer concentration to enhance protein expression. Autoinduction system optimization: the examination of glucose, lactose, and glycerol optimization within autoinduction systems to improve protein production. Osmolytes and osmoprotectants: an analysis of the use of osmolytes and osmoprotectants, such as sorbitol and glycine-betaine, to overcome with ease osmotic stress and enhance protein solubility. Ethanol additives: the impact of ethanol on E. coli physiology and its potential to improve recombinant protein expression. Cofactors and metabolic precursors: insights into the addition of cofactors, such as pyridoxal phosphate, riboflavin, thiamine, and pyridoxine, and the utilization of metabolic precursors to enhance the corresponding protein expression. This review highlights both the successful strategies and challenges in recombinant protein expression and provides insights into potential future research directions. Understanding and optimizing these factors is crucial for the efficient production of recombinant proteins for various applications in biotechnology. Furthermore, based on the analyzed data, we propose a straightforward scheme to optimize the additives in the cultivation medium.
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在大肠杆菌细胞质中表达可溶性重组蛋白的添加剂
在大肠杆菌中表达重组蛋白是分子生物学和生物技术中的一项基本技术。本综述全面概述了提高大肠杆菌中可溶性重组蛋白表达水平的各种添加剂。讨论包括五个主要方面。诱导剂优化:优化诱导剂浓度以提高蛋白质表达的策略。自动诱导系统优化:研究自动诱导系统中葡萄糖、乳糖和甘油的优化,以提高蛋白质产量。渗透剂和渗透保护剂:分析如何使用渗透剂和渗透保护剂(如山梨醇和甘氨酸-甜菜碱)来轻松克服渗透压力和提高蛋白质溶解度。乙醇添加剂:乙醇对大肠杆菌生理的影响及其改善重组蛋白表达的潜力。辅助因子和代谢前体:深入探讨添加辅助因子(如磷酸吡哆醛、核黄素、硫胺素和吡哆醇)和利用代谢前体提高相应蛋白质表达的方法。这篇综述强调了重组蛋白表达的成功策略和挑战,并对未来潜在的研究方向提出了见解。了解和优化这些因素对于高效生产用于生物技术各种应用的重组蛋白至关重要。此外,根据分析的数据,我们提出了优化培养基中添加剂的直接方案。
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